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Preparation method for plant source active component nano-scale membrane type vesicle

A technology for membrane vesicles and active ingredients, which is applied in the field of preparation of plant raw materials, can solve the problems of high material cost, limited material sources, restricting the industrial production and clinical application of exosomes, etc., and achieves the effects of low cost and abundant raw material sources.

Active Publication Date: 2014-01-01
居颂光
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Exosomes are obtained from sources such as cell culture supernatants. The source of materials is limited and the cost of materials is high. The content of exosomes obtained is very low, and the cost of purified exosomes is higher, which restricts the large-scale industrial production and clinical application of exosomes.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Cucumbers washed with clean water are squeezed out of juice by using a squeeze press in an environment of 20 degrees Celsius.

[0027] (2) The squeezed juice is filtered through 10-mesh, 100-mesh, and 600-mesh filters according to the mesh diameter from large to small (operated in an environment of 37 degrees Celsius), and the filtered liquid is collected.

[0028] (3) Centrifuge the collected liquid at 1000 rpm for 40 minutes, discard the precipitate, and collect the supernatant; centrifuge the supernatant at 2,000 rpm for 50 minutes, discard the precipitate, and collect the supernatant; centrifuge the supernatant at 6,000 rpm for 2 After 2 hours, the precipitate was discarded, and the supernatant was collected. The supernatant was centrifuged at 20,000 rpm for 3 hours to collect the precipitate, and the precipitate was resuspended with phosphate buffer to obtain a resuspension of membranous vesicles.

[0029] (4) The resuspension of membranous vesicles is h...

Embodiment 2

[0032] (1) The salvia miltiorrhiza washed with clean water is squeezed out by a squeeze press in an environment of 8 degrees Celsius.

[0033] (2) The squeezed juice is filtered through 800-mesh and 1200-mesh filters in turn (operated in an environment of 20 degrees Celsius), and the filtered liquid is collected.

[0034] (3) Centrifuge the collected liquid at 2000 rpm for 20 minutes, discard the precipitate, and collect the supernatant; centrifuge the supernatant at 4,000 rpm for 30 minutes, discard the precipitate, and collect the supernatant; centrifuge the supernatant at 10,000 rpm for 1 hour Discard the precipitate and collect the supernatant. The supernatant was centrifuged at 30,000 rpm for 1 hour to collect the precipitate, and the precipitate was resuspended with physiological saline to obtain a resuspension of membranous vesicles.

[0035] (4) The resuspension of membranous vesicles was homogenized with a nanoscale ultra-high pressure homogenizer to obtain nano...

Embodiment 3

[0038] (1) Squeeze out the juice from the garlic washed in water at 4 degrees Celsius.

[0039] (2) The squeezed juice is filtered through a 12500-mesh filter (operated in an environment of 10 degrees Celsius), and the filtered liquid is collected.

[0040] (3) Centrifuge the collected liquid at 3,000 rpm for 30 minutes, discard the precipitate, and collect the supernatant; centrifuge the supernatant at 6,000 rpm for 40 minutes, discard the precipitate, and collect the supernatant; centrifuge the supernatant at 30,000 rpm for 1 After 2 hours, the precipitate was discarded, and the supernatant was collected. The supernatant was centrifuged at 40,000 rpm for 1 hour to collect the precipitate, and the precipitate was resuspended with physiological saline to obtain a resuspension of membranous vesicles.

[0041] (4) The resuspension of membranous vesicles was homogenized with a shaker to obtain nano-membranous vesicles.

[0042] (5) Divide the samples, and the nanomembrane ...

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Abstract

The present invention provides a preparation method for a plant source active component nano-scale membrane type vesicle. The preparation method comprises the following steps: (1) extracting a plant raw material raw liquid; (2) extracting the supernatant or the concentrate; (3) precipitating and resuspending membrane type vesicle; and (4) carrying out a nano-scale homogenization treatment to obtain nanoparticles, wherein the nanoparticles are the plant source active component nano-scale membrane type vesicles. The present invention further provides a preparation method for the plant source active component nanoparticles with different densities. According to the present invention, the plant raw material is adopted to prepare the nano-scale membrane type vesicle, advantages of rich raw material source, low cost, mass production and the like are provided, and scalization vesicle utilization is achieved.

Description

technical field [0001] The invention relates to the preparation of plant raw materials, in particular to the preparation of a plant-derived active ingredient membranous vesicle. Background technique [0002] Membranous vesicles (exosomes) are cup-shaped small vesicles with a diameter of about 30-100 nm and a double-layer membrane structure that can be secreted by a variety of cells. Exosomes can transfer proteins, mRNA, microRNA and other active substances between cells, and participate in many important physiological and pathological processes, and their unique role has attracted more and more attention. [0003] At present, exosomes are mainly obtained from cell culture supernatant, tumor tissue exudate and other materials. The separation methods mainly include membrane filtration, ultracentrifugation, and sucrose density gradient centrifugation. Obtaining exosomes from sources such as cell culture supernatants has limited material sources and high material costs. The con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/00A61K9/51A23L1/29A23L33/00
Inventor 居颂光居颂文钱鸣宇
Owner 居颂光
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